饱和氢气生理盐水对大鼠肝移植缺血/再灌注损伤中自噬的调节作用  

Effects of hydrogen-rich saline on autophagy in rats underwent orthotopic liver transplantation

作　　者：施东婧 杜洪印[2] 喻文立[2] 吴莉[1] 盛明薇[2] 翁亦齐[2] 王永旺[2] 刘文娜[1] 王树森 

机构地区：[1]天津医科大学一中心临床学院,天津300070 [2]天津市第一中心医院麻醉科,天津300192 [3]国家卫生计生委危重病急救医学重点实验室,天津300192

出　　处：《实用器官移植电子杂志》2016年第1期18-22,1,共5页Practical Journal of Organ Transplantation(Electronic Version)

基　　金：天津市应用基础研究计划面上项目(05YFJMJC14800);天津市卫生行业重点攻关项目(13KG105)

摘　　要：目的研究饱和氢气生理盐水对大鼠原位肝移植缺血/再灌注损伤(IRI)中肝细胞自噬的调节作用。方法选取体重为220~250 g的健康雄性SD大鼠24只,采用随机数字表法将其分为3组(n=8):假手术组(Sham组)、原位肝移植组(OLT组)、饱和氢气生理盐水处理组(HS组)。Sham组仅进行开关腹操作。OLT组和HS组建立原位肝移植模型。HS组在肝下下腔开放即刻经肝下下腔静脉注射6 ml/kg含饱和氢气的生理盐水,OLT组则注射等量生理盐水。再灌注6 h及S组术毕6 h时取材,测定血清中丙氨酸转氨酶(ALT)和天冬氨酸转氨酶(AST)水平,观察肝组织形态学改变,检测氧化应激水平,应用蛋白质免疫印迹试验(Western Blot)检测微管相关蛋白轻链3(LC3-Ⅱ)和自噬相关因子Beclin-1蛋白表达,应用RT-PCR检测基因天冬氨酸特异性半胱氨酸蛋白酶3(caspase-3)和细胞色素C(Cyt-c)的m RNA表达水平。结果与Sham组相比,OLT组和HS组肝功能和病理损伤明显(P<0.05),MDA水平升高超氧化物歧化酶(SOD)活性降低显著〔MDA(nmol/mg):1.25±0.13、1.02±0.11比0.64±0.07,SOD(U/mg):135.78±10.92、158.81±12.09比204.13±17.75,均P<0.05〕,凋亡及自噬水平均升高(caspase:2.02±0.26、1.67±0.17比1.09±0.18,Cyt-c:1.96±0.19、1.64±0.18比0.99±0.17,均P<0.05)。HS组与OLT组相比,各项指标均下降〔MDA(nmol/mg):1.02±0.11比1.25±0.13,caspase-3(2^(-ΔΔCt)):1.67±0.17比2.02±0.26,Cyt-c(2^(-ΔΔCt)):1.64±0.18比1.96±0.19,均P<0.05〕。结论饱和氢气生理盐水可通过抑制自噬,减少细胞凋亡,有效缓解肝移植中IRI。Objective To investigate the effects of hydrogen-rich saline on autophagy in rats which undergoing orthotopic liver transplantation（OLT）. Methods Healthy male Sprague-Dawley（SD）rats, weighing 220-250 g, were divided into three groups（n = 8 each）： sham operated group（Sham group）; orthotopic liver transplantation group（OLT group）;orthotopic liver transplantation with hydrogen-rich saline treatment group（HS group）. Sham Group only underwent simple laparotomy. OLT were performed in OLT and HS group. In group HS, 6 ml/kg hydrogen-rich saline was injected through infrahepatic vena cava（IVC） after IVC reperfusion.All rats were sacrificed at 6 h after reperfusion. Hepatic function, oxidative stress（MDA and SOD） and pathological changes were detected. The expression of LC3- Ⅱ and Beclin-1 were detected by Western Blot. The expression of caspase-3 and Cyt-c m RNA were detected by RT-PCR. Results Compared with Sham group, the liver functions, the levels of oxidative stress, the expression of LC3- Ⅱ, Beclin-1、 caspase-3 m RNA and Cyt-c m RNA increased markedly in OLT and HS group（MDA（nmol/mg）：1.25±0.13、1.02±0.11 vs. 0.64±0.07,SOD（U/mg）： 135.78±10.92、158.81±12.09 vs. 204.13±17.75,caspase-3（2^（-ΔΔCt））：2.02±0.26、1.67±0.17 vs. 1.09±0.18,Cyt-C（2^（-ΔΔCt））：1.96±0.19、1.64±0.18 vs. 0.99±0.17,all P 〈0.05）. In HS group, the pathological changes,the levels of apoptosis and autophagy were significantly ameliorated compared to group OLT 〔MDA（nmol/mg ：1.02±0.11 vs. 1.25±0.13,caspase-3 ：1.67±0.17 vs. 2.02±0.26,Cyt-C ：1.64±0.18 vs. 1.96±0.19,all P 〈0.05）〕. Conclusion Hydrogen-rich saline can inhibit autophagy and attenuate ischemia reperfusion injury in rats undergoing OLT.

关 键 词：氢气 原位肝移植 缺血/再灌注损伤 自噬 

分 类 号：R657.3[医药卫生—外科学]